TRIzol Reagent
RNA isolation / purification Tissue - Mouse Adrenal glands
Protocol tips
| Protocol tips |
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| - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. |
Publication protocol
Total RNA was isolated from whole adrenal glands using TRIzol reagent (Life Technologies). Samples contained both adrenals from a mouse up to postnatal day (P) 15 or single adrenals after P15. For quantitative PCR (qPCR), groups of three or more samples were analyzed independently. The RNA pellet was dissolved in 11 μL water, and then 8 μL was incubated with oligo-deoxythymidine primers and SuperScript III reverse transcriptase (Life Technologies). First-strand cDNA template was mixed with 250 nM of test gene primers with Power SYBR master mix in a StepOne Plus real-time PCR system (Life Technologies). Relative RNA amounts for test genes were calculated using relative standard curves with Actb as a control, as an average of three or more samples from three mice and expressed relative to levels at P8.Full paper Login or join for free to view the full paper.
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Discussion
Discussion
5 years ago
Author:
Paul G. Macon
RNA isolation from tissue
How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?
Discussion
5 years ago
Author:
Aaron Stege
Problem in phase separation after using serum/plasma kit
I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?
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Manufacturer protocol
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